ERK5 Contributes to VEGF Alteration in Diabetic Retinopathy

Diabetic retinopathy is one of the most common causes of blindness in North America. Several signaling mechanisms are activated secondary to hyperglycemia in diabetes, leading to activation of vasoactive factors. We investigated a novel pathway, namely extracellular signal regulated kinase 5 (ERK5) mediated signaling, in modulating glucose-induced vascular endothelial growth factor (VEGF) expression. Human microvascular endothelial cells (HMVEC) were exposed to glucose. In parallel, retinal tissues from streptozotocin-induced diabetic rats were examined after 4 months of follow-up. In HMVECs, glucose caused initial activation followed by deactivation of ERK5 and its downstream mediators myocyte enhancing factor 2C (MEF2C) and Kruppel-like factor 2 (KLF2) mRNA expression. ERK5 inactivation further led to augmented VEGF mRNA expression. Furthermore, siRNA mediated ERK5 gene knockdown suppressed MEF2C and KLF2 expression and increased VEGF expression and angiogenesis. On the other hand, constitutively active MEK5, an activator of ERK5, increased ERK5 activation and ERK5 and KLF2 mRNA expression and attenuated basal- and glucose-induced VEGF mRNA expression. In the retina of diabetic rats, depletion of ERK5, KLF2 and upregulation of VEGF mRNA were demonstrated. These results indicated that ERK5 depletion contributes to glucose induced increased VEGF production and angiogenesis. Hence, ERK5 may be a putative therapeutic target to modulate VEGF expression in diabetic retinopathy

Position lifetime amd coincidence doppler broadening studies of graphene oxide - polyaniline nanocomposite

In this work, we report about the results of a detailed investigation comprising of measurements of positron lifetimes and coincidence Doppler broadening of the electron positron annihilation gamma ray spectra on graphene oxide – polyaniline nanocomposite samples prepared with different ratios of weight and at different temperatures. The two experimental techniques are capable of providing information respectively on the elec- tron density and momentum distribution at the specific sites of annihilation. This makes them effective in identifying the different types of defects present in the composite matrix. The magnitudes of the positron lifetimes were commensurate with the expectation of very large size defects in the form of vacancy clusters within the atomic composition of the composite material. The average defect size apparently reduced when the relative abundance of polyaniline increased that led to increased reduction of graphene oxide to graphene. The reduction also resulted in the shrinkage of the graphene oxide matrix and the free volume thereby released added to the overall defect concentration. The variation of the positron lifetime and its intensity with the temperature of synthesis suggested an optimum temperature suitable for the process. When you are citing the document, use the following link http://essuir.sumdu.edu.ua/handle/123456789/2062

Hydrogen Peroxide Inhibition of Bicupin Oxalate Oxidase

Oxalate oxidase is a manganese containing enzyme that catalyzes the oxidation of oxalate to carbon dioxide in a reaction that is coupled with the reduction of oxygen to hydrogen peroxide. Oxalate oxidase from Ceriporiopsis subvermispora (CsOxOx) is the first fungal and bicupin enzyme identified that catalyzes this reaction. Potential applications of oxalate oxidase for use in pancreatic cancer treatment, to prevent scaling in paper pulping, and in biofuel cells have highlighted the need to understand the extent of the hydrogen peroxide inhibition of the CsOxOx catalyzed oxidation of oxalate. We apply a membrane inlet mass spectrometry (MIMS) assay to directly measure initial rates of carbon dioxide formation and oxygen consumption in the presence and absence of hydrogen peroxide. This work demonstrates that hydrogen peroxide is both a reversible noncompetitive inhibitor of the CsOxOx catalyzed oxidation of oxalate and an irreversible inactivator. The build-up of the turnover-generated hydrogen peroxide product leads to the inactivation of the enzyme. The introduction of catalase to reaction mixtures protects the enzyme from inactivation allowing reactions to proceed to completion. Circular dichroism spectra indicate that no changes in global protein structure take place in the presence of hydrogen peroxide. Additionally, we show that the CsOxOx catalyzed reaction with the three carbon substrate mesoxalate consumes oxygen which is in contrast to previous proposals that it catalyzed a non-oxidative decarboxylation with this substrate

Effect of Binding Energies on the Encounter Desorption

The abundance of interstellar ice constituents is usually expressed with respect to the water ice because, in denser regions, a significant portion of the interstellar grain surface would be covered by water ice. The binding energy (BE) or adsorption energy of the interstellar species regulates the chemical complexity of the interstellar grain mantle. Due to the high abundance of water ice, the BE of surface species with the water is usually provided and widely used in astrochemical modeling. However, the hydrogen molecules would cover some part of the grain mantle in the denser and colder part of the interstellar medium. Even at around similar to 10 K, few atoms and simple molecules with lower adsorption energies can migrate through the surface. The BE of the surface species with H-2 substrate would be very different from that of a water substrate. However, adequate information regarding these differences is lacking. Here, we employ the quantum chemical calculation to provide the BE of 95 interstellar species with H-2 substrate. These are representative of the BEs of species to a H-2 overlayer on a grain surface. On average, we notice that the BE with the H-2 monomer substrate is almost ten times lower than the BE of these species reported earlier with the H2O c-tetramer configuration. The encounter desorption of H and H-2 was introduced [with E-D (H, H-2) = 45 K and E-D (H-2, H-2) = 23 K] to have a realistic estimation of the abundances of the surface species in the colder and denser region. Our quantum chemical calculations yield higher adsorption energy of H-2 than that of H [E-D (H, H-2) = 23-25 K and E-D (H-2, H-2) = 67-79 K]. We further implement an astrochemical model to study the effect of encounter desorption with the present realistic estimation. The encounter desorption of the N atom [calculations yield E-D (N, H-2) = 83 K] is introduced to study the differences with its inclusion

Securinine, a Myeloid Differentiation Agent with Therapeutic Potential for AML

As the defining feature of Acute Myeloid Leukemia (AML) is a maturation arrest, a highly desirable therapeutic strategy is to induce leukemic cell maturation. This therapeutic strategy has the potential of avoiding the significant side effects that occur with the traditional AML therapeutics. We identified a natural compound securinine, as a leukemia differentiation-inducing agent. Securinine is a plant-derived alkaloid that has previously been used clinically as a therapeutic for primarily neurological related diseases. Securinine induces monocytic differentiation of a wide range of myeloid leukemia cell lines as well as primary leukemic patient samples. Securinine\u27s clinical potential for AML can be seen from its ability to induce significant growth arrest in cell lines and patient samples as well as its activity in significantly impairing the growth of AML tumors in nude mice. In addition, securinine can synergize with currently employed agents such as ATRA and decitabine to induce differentiation. This study has revealed securinine induces differentiation through the activation of DNA damage signaling. Securinine is a promising new monocytic differentiation inducing agent for AML that has seen previous clinical use for non-related disorders

"Clumpiness" Mixing in Complex Networks

Three measures of clumpiness of complex networks are introduced. The measures quantify how most central nodes of a network are clumped together. The assortativity coefficient defined in a previous study measures a similar characteristic, but accounts only for the clumpiness of the central nodes that are directly connected to each other. The clumpiness coefficient defined in the present paper also takes into account the cases where central nodes are separated by a few links. The definition is based on the node degrees and the distances between pairs of nodes. The clumpiness coefficient together with the assortativity coefficient can define four classes of network. Numerical calculations demonstrate that the classification scheme successfully categorizes 30 real-world networks into the four classes: clumped assortative, clumped disassortative, loose assortative and loose disassortative networks. The clumpiness coefficient also differentiates the Erdos-Renyi model from the Barabasi-Albert model, which the assortativity coefficient could not differentiate. In addition, the bounds of the clumpiness coefficient as well as the relationships between the three measures of clumpiness are discussed.Comment: 47 pages, 11 figure

"Clumpiness" Mixing in Complex Networks

Three measures of clumpiness of complex networks are introduced. The measures quantify how most central nodes of a network are clumped together. The assortativity coefficient defined in a previous study measures a similar characteristic, but accounts only for the clumpiness of the central nodes that are directly connected to each other. The clumpiness coefficient defined in the present paper also takes into account the cases where central nodes are separated by a few links. The definition is based on the node degrees and the distances between pairs of nodes. The clumpiness coefficient together with the assortativity coefficient can define four classes of network. Numerical calculations demonstrate that the classification scheme successfully categorizes 30 real-world networks into the four classes: clumped assortative, clumped disassortative, loose assortative and loose disassortative networks. The clumpiness coefficient also differentiates the Erdos-Renyi model from the Barabasi-Albert model, which the assortativity coefficient could not differentiate. In addition, the bounds of the clumpiness coefficient as well as the relationships between the three measures of clumpiness are discussed.Comment: 47 pages, 11 figure

Identification of a Novel Synaptic Protein, TMTC3, Involved in Periventricular Nodular Heterotopia with Intellectual Disability and Epilepsy

Defects in neuronal migration cause brain malformations, which are associated with intellectual disability (ID) and epilepsy. Using exome sequencing, we identified compound heterozygous variants (p.Arg71His and p. Leu729ThrfsTer6) in TMTC3, encoding transmembrane and tetratricopeptide repeat containing 3, in four siblings with nocturnal seizures and ID. Three of the four siblings have periventricular nodular heterotopia (PVNH), a common brain malformation caused by failure of neurons to migrate from the ventricular zone to the cortex. Expression analysis using patient-derived cells confirmed reduced TMTC3 transcript levels and loss of the TMTC3 protein compared to parental and control cells. As TMTC3 function is currently unexplored in the brain, we gathered support for a neurobiological role for TMTC3 by generating flies with post-mitotic neuron-specific knockdown of the highly conserved Drosophila melanogaster TMTC3 ortholog, CG4050/tmtc3. Neuron-specific knockdown of tmtc3 in flies resulted in increased susceptibility to induced seizures. Importantly, this phenotype was rescued by neuron-specific expression of human TMTC3, suggesting a role for TMTC3 in seizure biology. In addition, we observed co-localization of TMTC3 in the rat brain with vesicular GABA transporter (VGAT), a presynaptic marker for inhibitory synapses. TMTC3 is localized at VGAT positive pre-synaptic terminals and boutons in the rat hypothalamus and piriform cortex, suggesting a role for TMTC3 in the regulation of GABAergic inhibitory synapses. TMTC3 did not co-localize with Vglut2, a presynaptic marker for excitatory neurons. Our data identified TMTC3 as a synaptic protein that is involved in PVNH with ID and epilepsy, in addition to its previously described association with cobblestone lissencephaly

Diagnostic Utility of Genome-wide DNA Methylation Testing in Genetically Unsolved Individuals with Suspected Hereditary Conditions.

Conventional genetic testing of individuals with neurodevelopmental presentations and congenital anomalies (ND/CAs), i.e., the analysis of sequence and copy number variants, leaves a substantial proportion of them unexplained. Some of these cases have been shown to result from DNA methylation defects at a single locus (epi-variants), while others can exhibit syndrome-specific DNA methylation changes across multiple loci (epi-signatures). Here, we investigate the clinical diagnostic utility of genome-wide DNA methylation analysis of peripheral blood in unresolved ND/CAs. We generate a computational model enabling concurrent detection of 14 syndromes using DNA methylation data with full accuracy. We demonstrate the ability of this model in resolving 67 individuals with uncertain clinical diagnoses, some of whom had variants of unknown clinical significance (VUS) in the related genes. We show that the provisional diagnoses can be ruled out in many of the case subjects, some of whom are shown by our model to have other diseases initially not considered. By applying this model to a cohort of 965 ND/CA-affected subjects without a previous diagnostic assumption and a separate assessment of rare epi-variants in this cohort, we identify 15 case subjects with syndromic Mendelian disorders, 12 case subjects with imprinting and trinucleotide repeat expansion disorders, as well as 106 case subjects with rare epi-variants, a portion of which involved genes clinically or functionally linked to the subjects\u27 phenotypes. This study demonstrates that genomic DNA methylation analysis can facilitate the molecular diagnosis of unresolved clinical cases and highlights the potential value of epigenomic testing in the routine clinical assessment of ND/CAs